RESUMO
T lymphocytes play a critical role in antitumor immunity, but their exhaustion poses a significant challenge for immune evasion by malignant cells. Circular RNAs (circRNAs), characterized by their covalently closed looped structure, have emerged as pivotal regulators within the neoplastic landscape. Recent studies have highlighted their multifaceted roles in cellular processes, including gene expression modulation and protein function regulation, which are often disrupted in cancer. In this review, we systematically explore the intricate interplay between circRNAs and T cell modulation within the tumor microenvironment. By dissecting the regulatory mechanisms through which circRNAs impact T cell exhaustion, we aim to uncover pathways crucial for immune evasion and T cell dysfunction. These insights can inform innovative immunotherapeutic strategies targeting circRNA-mediated molecular pathways. Additionally, we discuss the translational potential of circRNAs as biomarkers for therapeutic response prediction and as intervention targets. Our comprehensive analysis aims to enhance the understanding of immune evasion dynamics in the tumor microenvironment by facilitating the development of precision immunotherapy.
Assuntos
Neoplasias , RNA Circular , Linfócitos T , Microambiente Tumoral , Humanos , RNA Circular/genética , Neoplasias/imunologia , Neoplasias/genética , Neoplasias/terapia , Neoplasias/patologia , Neoplasias/metabolismo , Microambiente Tumoral/imunologia , Microambiente Tumoral/genética , Linfócitos T/imunologia , Linfócitos T/metabolismo , Regulação Neoplásica da Expressão Gênica , Imunoterapia/métodos , Animais , Biomarcadores Tumorais/genéticaRESUMO
Gastric cancer (GC) is the third leading cause of cancer-related death worldwide. Due to the lack of early symptoms, GC is often diagnosed at an advanced stage when treatment options are limited. There is an urgent need to identify biomarkers for early detection, prognosis evaluation, and targeted treatment of GC. Studies have shown that Src kinase-associated phosphoprotein 1 (SKAP1) promotes cell proliferation and invasion and is associated with poor prognosis in colorectal cancer, malignant fibrous histiocytoma, and breast cancer. However, the role and mechanism of SKAP1 in GC are unclear. Here, analyses of multiple databases and experiments revealed that SKAP1 expression was higher in GC than in adjacent normal tissues. The Cancer Genome Atlas data showed that high SKAP1 expression was associated with poor GC prognosis. SKAP1 expression was higher in GC than in normal gastric epithelial cells. SKAP1 silencing reduced the proliferation, migration and invasion of the GC cell lines MKN45 and HGC27. Rescue experiments suggest that SKAP1 may promote GC progression by activating JAK1/PI3K/AKT signaling and regulating GC cell proliferation, invasion, migration, and other functions. Bioinformatics analysis revealed that SKAP1 was associated with immune cell infiltration and checkpoint expression in GC. High SKAP1 expression was associated with poorer immunotherapy outcomes, suggesting its potential as a predictive biomarker of GC immunotherapy efficacy. In summary, SKAP1 is overexpressed in GC, where it promotes cell proliferation, invasion and migration and is associated with poor prognosis and poor immunotherapy outcomes. SKAP1 may represent a biomarker and therapeutic target in GC and regulates cellular functions through JAK1/PI3K/AKT signaling.
Assuntos
Neoplasias Gástricas , Humanos , Neoplasias Gástricas/genética , Neoplasias Gástricas/terapia , Neoplasias Gástricas/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Movimento Celular/genética , Proliferação de Células/genética , Biomarcadores , Regulação Neoplásica da Expressão Gênica , Linhagem Celular Tumoral , Fosfoproteínas/metabolismoRESUMO
Long non-coding RNAs (LncRNAs), lacking protein-coding function, modulate immune function by regulating the expression of genes or the function of protein molecules. They participate in epigenetic regulation, interfere with downstream gene transcription acting as a molecular sponge to affect miRNA function, and can combine with proteins to form nucleic acid protein complexes that affect protein function or cell location to regulate genes and regulate immune function. LncRNAs are differentially expressed in immune cells. They affect the maturity, differentiation and activation of immune cells and regulate cytokine release and immune phenotype. They are closely related to immune tolerance and cell migration. Dendritic cells (DCs) are important immune cells with the most robust antigen-presenting function, and have irreplaceable roles in human innate immunity and adaptive immunity. Emerging evidence over the past few years has suggested that LncRNAs influence the differentiation and maturation of DCs and serve as a critical role in the cell phenotype and immune function of DCs. To further understand the role of LncRNAs in the occurrence and development of DC-related diseases, we elaborated the role of LncRNAs in DC immune function, including antigen presentation, T cell activation and proliferation, DC migration. Furthermore, we summarized the impact of pathological factors (tumors, inflammation, autoimmune disease, viral infection) and physiological factors (e.g., age) on the LncRNAs in DCs, and how the changed LncRNAs altered the function and behavior of DCs resulting from the intervention. We hope this review give us have a better understanding of multiple effects of LncRNA on cell function in DCs.
Assuntos
MicroRNAs , RNA Longo não Codificante , Humanos , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Células Dendríticas , Epigênese Genética , Diferenciação Celular , MicroRNAs/metabolismoRESUMO
Immune checkpoint inhibitors (ICIs), including anti-programmed cell death-1/anti-programmed cell death ligand-1 (anti-PD-1/PD-L1) therapy, have elicited impressive clinical outcomes in several malignancies. This is regarded as a pivotal breakthrough in cancer treatment. However, a vast majority of colorectal cancer (CRC) cases are microsatellite stable (MSS) and respond poorly to anti-PD-1/PD-L1 immunotherapies. Since ICIs serve as rescuers for immune cell-mediated cancer cell elimination, the limited efficacy of anti-PD-1/PD-L1 treatments may be attributed to the privileged tumor microenvironment (TME), which is characterized by unavailable immunosurveillance. Thus, it is essential to modify the pre-existing disordered immune system prior to the application of an anti-PD-1/PD-L1 therapy. In this review, to overcome unsatisfactory immunotherapy in CRC patients with MSS, we discussed various combination therapies based on TME reconstruction for improving the susceptibility to anti-PD-1/PD-L1 treatment.
Assuntos
Antígeno B7-H1 , Neoplasias Colorretais , Neoplasias Colorretais/tratamento farmacológico , Neoplasias Colorretais/patologia , Humanos , Fatores Imunológicos/uso terapêutico , Imunoterapia , Instabilidade de Microssatélites , Microambiente TumoralRESUMO
Dendritic cell (DC) based immunotherapy is a promising approach to clinical cancer treatment. miRNAs are a class of small non-coding RNA molecules that bind to RNAs to mediate multiple events which are important in diverse biological processes. miRNA mimics and antagomirs may be potent agents to enhance DC-based immunotherapy against cancers. miRNA array analysis was used to identify a representative miR-5119 potentially regulating PD-L1 in DCs. We evaluated levels of ligands of immune cell inhibitory receptors (IRs) and miR-5119 in DCs from immunocompetent mouse breast tumor-bearing mice, and examined the molecular targets of miR-5119. We report that miRNA-5119 was downregulated in spleen DCs from mouse breast cancer-bearing mice. In silico analysis and qPCR data showed that miRNA-5119 targeted mRNAs encoding multiple negative immune regulatory molecules, including ligands of IRs such as PD-L1 and IDO2. DCs engineered to express a miR-5119 mimic downregulated PD-L1 and prevented T cell exhaustion in mice with breast cancer homografts. Moreover, miR-5119 mimic-engineered DCs effectively restored function to exhausted CD8+ T cells in vitro and in vivo, resulting in robust anti-tumor cell immune response, upregulated cytokine production, reduced T cell apoptosis, and exhaustion. Treatment of 4T1 breast tumor-bearing mice with miR-5119 mimic-engineered DC vaccine reduced T cell exhaustion and suppressed mouse breast tumor homograft growth. This study provides evidence supporting a novel therapeutic approach using miRNA-5119 mimic-engineered DC vaccines to regulate inhibitory receptors and enhance anti-tumor immune response in a mouse model of breast cancer. miRNA/DC-based immunotherapy has potential for advancement to the clinic as a new strategy for DC-based anti-breast cancer immunotherapy.
Assuntos
Neoplasias da Mama/tratamento farmacológico , Vacinas Anticâncer/imunologia , Células Dendríticas/imunologia , Imunoterapia/métodos , MicroRNAs/metabolismo , Animais , Neoplasias da Mama/imunologia , Linhagem Celular Tumoral , Modelos Animais de Doenças , Feminino , Humanos , Camundongos , TransfecçãoRESUMO
Abnormally high expression of glial cell line-derived neurotrophic factor (GDNF) derived from glioma cells has essential impacts on gliomagenesis and development, but the molecular basis underlying increased GDNF expression in glioma cells remain unclear. This work aimed to study the molecular mechanisms that may explain the accumulation of GDNF in glioma. Firstly, we observed that cAMP response element-binding protein (CREB), known as an important transcription factor for binding of GDNF promoter region, was highly expressed with an apparent accumulation into the nucleus of glioma cells, which may contribute to the transcription of GDNF. Secondly, CUE domain-containing protein 2 (CUEDC2), a ubiquitin-regulated protein, could increase the amount of binding between the E3 ligase tripartite motif-containing 21 (TRIM21) and CREB and affect the CREB level. Like our previous study, it showed that there was a significantly down-regulation of CUEDC2 in glioma. Finally, our data suggest that GDNF expression is indirectly regulated by transcription factor ubiquitination. Indeed, down-regulation of CUEDC2, decreased the ubiquitination and degradation of CREB, which was associated to high levels of GDNF. Furthermore, abundant CREB involved in the binding to the GDNF promoter region contributes to GDNF high expression in glioma cells. Collectively, it was verified the GDNF expression was affected by CREB ubiquitination regulated by CUEDC2 level.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Glioma/metabolismo , Ubiquitinação/fisiologia , Linhagem Celular Tumoral , Regulação para Baixo , Regulação Neoplásica da Expressão Gênica/fisiologia , Glioma/genética , HumanosRESUMO
OBJECTIVE: To investigate the preventive effect of local injection of botulinum toxin type A (BTX-A) against iatrogenic anterior urethral stricture (IAUS) in New Zealand male rabbits. METHODS: Sixteen adult New Zealand male rabbits were randomly divided into an experimental and a control group of equal number. The model of IAUS was established in the rabbits by incision of the ventral urethra of the penile segment and electrocoagulation of the urethral mucosa. The rabbits of the experimental group were injected with 10 U of BTX-A solution and those of the control group with 1.0 ml of normal saline into the electrocoagulated submucosa of the urethra. At 30 days after modeling, retrograde urethrography was performed and the scarred urethral tissue harvested for HE and Masson staining. RESULTS: No systemic symptoms of botulinum toxin poisoning were observed in either group of the rabbits. Retrograde urethrography showed statistically significant differences between the control and experimental groups in the diameter of the urethra (ï¼»0.15 ± 0.08ï¼½ vs ï¼»0.50 ± 0.23ï¼½ cm, P < 0.05) and the stenosis ratio (3.68 ± 1.22 vs 1.29 ± 0.15, P < 0.05). Urethrography revealed obvious hyperplasia and contracture of urethral scar with high bulge and narrow urethral cavity, while HE and Masson staining exhibited absence or discontinuity of the urethral epithelium, submucosal fibrosis with infiltration of a large number of fibroblasts, and hyperplasia and thickening of collagen fibers (blue) in the control group. All the changes above were slight and the urethral epithelium was continuous in the experimental group. CONCLUSIONS: Local injection of BTX-A reduced the incidence of iatrogenic anterior urethral stricture after iatrogenic acute heat injury in New Zealand male rabbits.
Assuntos
Toxinas Botulínicas Tipo A/administração & dosagem , Estreitamento Uretral , Animais , Doença Iatrogênica/prevenção & controle , Injeções , Masculino , Coelhos , Distribuição Aleatória , Uretra , Estreitamento Uretral/prevenção & controleRESUMO
Decrease of chloride concentration contributes to cardiovascular diseases, however, whether decrease of chloride concentration is involved in platelet activation remains elusive. In the present study, we found that ACI patients had lower serum chloride which would be rescued after Aspirin administration. ADP induced chloride concentration reduction in platelets. Blockade of chloride channel prevented ADP-induced platelet adhesion, activation and aggregation, however, decreasing the extracellular chloride concentration promoted ADP-induced platelet adhesion and activation. Decrease of the extracellular chloride concentration facilitated the inactivation of Src family kinase Lyn, which was not involved in PI3K/Akt phosphorylation. Nevertheless, low chloride concentration promoted the production of platelet cytosol Gαi2 subunit. This subunit prevents AC from converting ATP into cAMP, which therefore, inhibited the phosphorylation of PKA to promote platelet activation. In conclusion, decreased intracellular chloride promotes ADP induced platelet activation through the Gαi2/cAMP/PKA pathway instead of the Lyn/PI3K/Akt signal pathway.
Assuntos
Difosfato de Adenosina/metabolismo , Cloretos/farmacologia , Proteínas Quinases Dependentes de AMP Cíclico/antagonistas & inibidores , AMP Cíclico/antagonistas & inibidores , Ativação Plaquetária/efeitos dos fármacos , Inibidores da Agregação Plaquetária/farmacologia , Transdução de Sinais/efeitos dos fármacos , Animais , Cloretos/sangue , AMP Cíclico/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Humanos , Camundongos , Fosfatidilinositol 3-Quinases/metabolismo , Inibidores da Agregação Plaquetária/sangue , Proteínas Proto-Oncogênicas c-akt/metabolismo , Quinases da Família src/metabolismoRESUMO
BACKGROUND/AIMS: Glial cell line-derived neurotrophic factor (GDNF) is an important factor promoting invasive glioma growth. This study was performed to reveal a unique mechanism of glioma cell proliferation and migration. METHODS: Human U251 glioma cells were used to screen the optimal GDNF concentration and treatment time to stimulate proliferation and migration. MicroRNA (MiRNA) expression profiles were detected by microarray and confirmed by real-time polymerase chain reaction (PCR). The target genes of differentially expressed miRNAs were predicted by miRWalk, and those targeted by multiple miRNAs were screened with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. A regulatory miRNA network was constructed using ingenuity pathway analysis (IPA). Target gene expression of differentially expressed miRNAs was examined by real-time PCR or mRNA microarray. RESULTS: The results show that 50 ng/mL GDNF for 24 h significantly promotes U251 glioma cell proliferation and migration (P < 0.05). Seven miRNAs (hsa-miR-194-5p, hsa-miR-152-3p, hsa-miR-205-5p, hsa-miR-629-5p, hsa-miR-3609, hsa-miR-183-5p, and hsa-miR-487b-3p) were significantly up-regulated after GDNF treatment (P < 0.05). These miRNAs are primarily involved in signal transduction, cell adhesion and cell cycle through mitogen-activated protein kinase (MAPK) signaling, focal adhesion and glioma signal pathways. Five of these miRNAs (hsa-miR-194-5p, hsa-miR-152-3p, hsa-miR-205-5p, hsa-miR-183-5p, and hsa-miR-487b-3p) co-regulate TP53 and Akt. mRNA expression levels of four genes co-targeted by two or more up-regulated miRNAs were significantly decreased after GDNF treatment (P < 0.05). CONCLUSION: GDNF treatment of U251 glioma cells significantly increased the expression of seven miRNAs involved in cell adhesion and the cell cycle.
Assuntos
Proliferação de Células/efeitos dos fármacos , Fator Neurotrófico Derivado de Linhagem de Célula Glial/farmacologia , MicroRNAs/metabolismo , Adesão Celular/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Análise por Conglomerados , Glioma/metabolismo , Glioma/patologia , Humanos , MicroRNAs/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais/efeitos dos fármacos , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Regulação para Cima/efeitos dos fármacosRESUMO
Inflammation plays a key role in the pathogenesis of many retinal degenerative diseases related with photoreceptor dysfunction/degeneration. However the involvement of photoreceptor cells in inflammatory reactions is largely unknown as they are not considered as inflammatory cells. In this study, we assessed whether photoreceptor cells can produce CCL2 and CXCL10, two important players in inflammation during endoplasmic reticulum (ER) stress. After photoreceptor 661 W cells were treated with ER stress inducer thapsigargin (TG), induction of ER stress increased CXCL10 and CCL2 expression at both mRNA and protein levels, which was significantly blocked by an ER stress blocker 4-phenylbutyrate. ER stress contains three pathways: PERK, ATF6 and IRE1α. Knockdown of PERK attenuated TG-induced CXCL10 and CCL2 mRNA expression, associated with significant decreases in phosphorylation of NF-κB RelA and STAT3. In contrast to PERK, knockdown of XBP1, which is activated by IRE1α-mediated splicing, robustly enhanced TG-induced CXCL10 and CCL2 expression and phosphorylation of NF-κB RelA and STAT3. Blockade of NF-κB or STAT3 markedly diminished TG-induced CXCL10 and CCL2 expression. The specific roles of PERK and XBP1 in CXCL10 and CCL2 expression were further investigated by treating photoreceptor cells with advanced glycation end products (AGE) and high glucose (HG), two of the major contributors to diabetic complications. Similarly, AGE and HG induced CXCL10 and CCL2 expression in which PERK was a positive regulator while XBP1 was a negative regulator. These studies suggest that photoreceptors may be involved in retinal inflammation by expressing chemokines CXCL10 and CCL2. PERK and IRE1α/XBP1 in the unfolded protein response differentially regulate the expression of CXCL10 and CCL2 likely through modulation of ER stress-induced NF-κB RelA and STAT3 activation.
Assuntos
Quimiocina CCL2/genética , Quimiocina CXCL10/genética , Estresse do Retículo Endoplasmático/genética , Regulação da Expressão Gênica , Células Fotorreceptoras de Vertebrados/metabolismo , Proteína 1 de Ligação a X-Box/metabolismo , eIF-2 Quinase/metabolismo , Animais , Western Blotting , Sobrevivência Celular , Células Cultivadas , Quimiocina CCL2/biossíntese , Quimiocina CXCL10/biossíntese , Citocinas , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Camundongos , Camundongos Knockout , Células Fotorreceptoras de Vertebrados/citologia , Reação em Cadeia da Polimerase , RNA/genética , Degeneração Retiniana/genética , Degeneração Retiniana/metabolismo , Degeneração Retiniana/patologiaRESUMO
In recent years, the role of capsaicin in cancer prevention and treatment has gained people's attention. However, the mechanism of anti-glioma cells by capsaicin has not been elucidated. Here, we discuss the mechanism of capsaicin in U251 cells. Cell viability was detected by MTT and extracellular LDH measurements, while immunofluorescence was performed to measure changes of LC3 in U251 cells. The expressions of LC3II, Puma-α, Beclin1, P62, Procaspase-3, and P53 were observed by immunoblotting. The cell viability decreased and the punctate patterns of LC3 in U251 cells were observed after Capsaicin treatment. Meanwhile, the expressions of Beclin1, P62, and Puma-α increased. After using 3-MA, the expressions of Beclin1 and Procaspase-3 were reduced while those of P53 and Puma-α increased. The expression of LC3II was increased after Pifithrin-α treatment. Therefore, we believed that capsaicin could induce apoptosis in U251 cells, and the inhibition of autophagy could contribute to apoptosis.
Assuntos
Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Capsaicina/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Benzotiazóis/farmacologia , Caspase 3/metabolismo , Linhagem Celular Tumoral , Glioma/tratamento farmacológico , Humanos , Transdução de Sinais/efeitos dos fármacos , Tolueno/análogos & derivados , Tolueno/farmacologiaRESUMO
Lipoxins (LXs) display unique pro-resolving and anti-inflammatory functions in a variety of inflammatory conditions. The present study was undertaken to investigate the effects of BML-111 (5(S),6(R),7-trihydroxyheptanoic acid methyl ester), the agonist of lipoxin A4 receptor, in a model of Lipopolysaccharides (LPS) and d-Galactosamine (d-GalN) induced acute liver injury, and to explore the mechanisms. Histopathological analyses were carried out to quantify liver injury degree. The activities of myeloperoxidase (MPO) were examined to evaluate the levels of neutrophil infiltration. The activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in serum were detected to evaluate the functions of the liver. The amounts of tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), and interleukin-1ß (IL-1ß) were measured using enzyme-linked immunosorbent assay (ELISA), and the expression levels of transforming growth factor-ß1(TGF-ß1) and cyclooxygenase-2 (COX-2) were examined using Western blotting. The antioxidant capacity, the activities of inducible nitric oxide synthase (iNOS), the contents of malondialdehyde (MDA) and nitric oxide (NO) were analyzed with the kits via biochemical analysis. We established the model of acute liver injury with lipopolysaccharide and d-Galactosamine (LPS/d-GalN): (1) histopathological results and MPO activities, with the activities of AST and ALT in serum, consistently demonstrated LPS and d-GalN challenge could cause severe liver damage, but BML-111 could prevent pathological changes, inhibit neutrophil infiltration, and improve the hepatic function; (2) LPS/d-GalN increased TNF-α, IL-1ß, COX-2, and IL-10, while decreasing TGF-ß1. However, BML-111 could repress LPS/d-GalN -induced TNF-α, IL-1ß and COX-2, meanwhile increasing the expression levels of TGF-ß1 and IL-10; (3) LPS/d-GalN inhibited the activities of superoxide dismutase (SOD), catalase (CAT), total antioxidant capacity (T-AOC), and hydroxyl radical-scavenging ability, simultaneously increasing the levels of MDA and NO, so also the activity of iNOS. Otherwise, BML-111 could reverse all the phenomena. In a word, BML-111 played a protective role in acute liver injury induced by LPS and d-GalN in rats, through improving antioxidant capacity and regulating the balance of inflammatory cytokines.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas/tratamento farmacológico , Galactosamina/toxicidade , Ácidos Heptanoicos/farmacologia , Lipopolissacarídeos/toxicidade , Substâncias Protetoras/farmacologia , Doença Aguda , Animais , Antioxidantes/metabolismo , Western Blotting , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Citocinas/metabolismo , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
The engulfment and cell motility (ELMOs) family of proteins plays a crucial role in tumor cell migration and invasion. However, the function of ELMO3 is poorly defined. To elucidate its role in the development and progression of colorectal cancer (CRC), we examined the expression of ELMO3 in 45 cases of paired CRC tumor tissues and adjacent normal tissues. Furthermore, we assessed the effect of the knockdown of ELMO3 on cell proliferation, cell cycle, migration, invasion and F-actin polymerization in HCT116 cells. The result shows that the expression of ELMO3 in CRC tissues was significantly increased in comparison to the adjacent normal colorectal tissues. Moreover, this overexpression was associated with tumor size (p = 0.007), tumor differentiation (p = 0.001), depth of invasion (p = 0.009), lymph node metastasis (p = 0.003), distant metastasis (p = 0.013) and tumor, node, metastasis (TNM)-based classification (p = 0.000). In in vitro experiments, the silencing of ELMO3 inhibited cell proliferation, invasion, metastasis, and F-actin polymerization, and induced Gap 1 (G1) phase cell cycle arrest. Our study demonstrates that ELMO3 is involved in the processes of growth, invasion and metastasis of CRC, and could be used a potential molecular diagnostic tool or therapy target of CRC.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/genética , Movimento Celular/genética , Proliferação de Células/genética , Neoplasias Colorretais/genética , Proteínas do Citoesqueleto/genética , Pontos de Checagem da Fase G1 do Ciclo Celular/genética , Actinas/metabolismo , Linhagem Celular Tumoral , Neoplasias Colorretais/patologia , Feminino , Células HCT116 , Humanos , Metástase Linfática/genética , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica/genética , Interferência de RNA , RNA Interferente Pequeno/genéticaRESUMO
Objective: To compare the incidences of Y chromosome microdeletion between patients with azoospermia or severe oligozoospermia with varicocele( VC) and those without VC and investigate the etiopathogenisis of their infertility. Methods: We included 137 VC patients in group A(70 with azoospermia as group A1 and 67 with severe oligozoospermia as group A2),135 non-VC patients in group B(69 with azoospermia as group B1 and 66 with severe oligozoospermia as group B2),and 30 normal fertile men as controls in group C. We detected Y chromosome microdeletion in different groups using multiplex PCR. Results: Y chromosome microdeletion was detected in 23(16. 8%) of the patients in group A, another 23(17. 0%) in group B,and 0 in group C. The rates of Y chromosome microdeletion were 22. 9% in group A1,10. 4% in group A2,20. 3% in group B1,and 13. 6% in group B2,and the microdeletion rate in the patients with severe oligozoospermia( groups A1 and B1) was 23. 3%(31 /133). No statistically significant difference was found between groups A and B( P > 0. 05). Conclusion: There are no significant differences in the rate of Y chromosome microdeletion between varicocele and non-varicocele patients with azoospermia or severe oligozoospermia, and Y chromosome microdeletion is one of the causes of azoospermia and severe oligozoospermia with varicocele.
Assuntos
Azoospermia/genética , Infertilidade Masculina/genética , Oligospermia/genética , Transtornos do Cromossomo Sexual no Desenvolvimento Sexual/genética , Varicocele/complicações , Estudos de Casos e Controles , Deleção Cromossômica , Cromossomos Humanos Y/genética , Humanos , Incidência , Masculino , Aberrações dos Cromossomos SexuaisRESUMO
Abnormally high transcription of the glial cell-line derived neurotrophic factor (gdnf) gene in glioma cells is related to the hyperacetylation of histone H3 lysine 9 (H3K9) in its promoter region II, but the mechanism remains unclear. There are three consecutive putative binding sites for the transcription factor early growth response protein 1(Egr-1) in promoter region II of the gdnf gene, and Egr-1 participates in gdnf gene transcription activation. Here we show that the acetylation level of H3K9 at Egr-1 binding sites in gdnf gene promoter region II in rat C6 astroglioma cells was significantly higher than that in normal astrocytes, and the binding capacity was also significantly higher. In C6 astroglioma cells, gdnf gene transcription significantly decreased after Egr-1 knock-down. In addition, the deletion or mutation of the Egr-1 binding site also significantly down-regulated the activity of promoter region II of this gene in vitro. When curcumin decreased the acetylation level of H3K9 at the Egr-1 binding site, the binding of Egr-1 to promoter region II and GDNF mRNA levels significantly decreased. In contrast, trichostatin A treatment significantly increased H3K9 acetylation at the Egr-1 binding site, which significantly increased both the binding of Egr-1 with promoter region II and GDNF mRNA levels. In this context, knocking down Egr-1 significantly reduced the elevation in gdnf gene transcription. Collectively, our results demonstrate that the hyperacetylation of H3K9 at Egr-1 binding sites in promoter region II of the gdnf gene can up-regulate the binding of Egr-1 to increase gdnf gene transcription in glioma cells.
Assuntos
Proteína 1 de Resposta de Crescimento Precoce/fisiologia , Fator Neurotrófico Derivado de Linhagem de Célula Glial/genética , Glioma/genética , Glioma/metabolismo , Histona Acetiltransferases/metabolismo , Histonas/metabolismo , Acetilação , Animais , Sítios de Ligação , Células Cultivadas , Regulação Neoplásica da Expressão Gênica , Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Regiões Promotoras Genéticas , RatosRESUMO
OBJECTIVE: To investigate the relationship between the level of di-2-ethylhexyl phthalate (DEHP) and idiopathic oligoasthenospermia by measuring the content of DEHP in the semen samples of different subjects. METHODS: We obtained semen samples from 100 infertile men with idiopathic oligoasthenospermia, 50 working all the year round in the plastic greenhouse (group A) and the other 50 constantly dining from plastic meal boxes (group B). We also enrolled 50 normal male volunteers as controls (group C). We conducted semen analyses using a computer-assisted sperm analyzer, measured the DEHP concentration by reversed-phase high-pressure liquid chromatography, and subjected the data to statistic processing by t-test and correlation analysis. RESULTS: The mean concentrations of DEHP in the seminal plasma were (0.72 +/- 0.48), (0.71 +/- 0.49) and (0.21 +/- 0.18) mg/L in groups A, B and C, respectively, significantly higher in A and B than in C (both P < 0.05). The DEHP concentration was negatively correlated with sperm motility (P < 0.05). CONCLUSION: The DEHP level in the seminal plasma is higher in infertile men frequently exposed to plastic products than in normal males and excessive DEHP may be one of the important factors of idiopathic male infertility.
Assuntos
Dietilexilftalato/efeitos adversos , Oligospermia/etiologia , Adulto , Estudos de Casos e Controles , Humanos , Masculino , Plásticos/efeitos adversos , Sêmen/químicaRESUMO
BACKGROUND: Cuproptosis is a type of cell death characterized by excessive copper-lipid reactions in the tricarboxylic acid cycle, resulting in protein toxicity stress and cell death. Although known as a cuproptosis inhibitor through CRISPR-Cas9 screening, the role of cyclin-dependent kinase inhibitor 2A (CDKN2A) in cuproptosis resistance and its connection to tumor development remains unclear. METHODS: In this study, we combined single-cell sequencing, spatial transcriptomics, pathological image analysis, TCGA multi-omics analysis and in vitro experimental validation to comprehensively investigate CDKN2A distribution, expression, epigenetic modification, regulation and genomic features in colorectal cancer cells. We further explored the associations between CDKN2A and cellular pathway, immune infiltration and spatial signal communication. RESULTS: Our findings showed an increasing trend in cuproptosis in the trajectory of tumor progression, accompanied by an upward trend of CDKN2A. CDKN2A underwent transcriptional activation by MEF2D and via the SNHG7/miR-133b axis, upregulating glycolysis, copper metabolism and copper ion efflux. CDKN2A likely drives epithelial-mesenchymal transition (EMT) and progression by activating Wnt signaling. CDKN2A is associated with high genomic instability and sensitivity to radiation and chemotherapy. Tumor regions expressing CDKN2A exhibit distinctive SPP1+ tumor-associated macrophage (TAM) infiltration and MMP7 enrichment, along with unique signaling crosstalk with adjacent areas. CONCLUSIONS: CDKN2A mediates cuproptosis resistance through regulating glycolysis and copper homeostasis, accompanied by a malignant phenotype and pro-tumor niche. Radiation and chemotherapy are expected to potentially serve as therapeutic approaches for cuproptosis-resistant colorectal cancer with high CDKN2A expression.
Assuntos
Neoplasias Colorretais , Inibidor p16 de Quinase Dependente de Ciclina , Progressão da Doença , Humanos , Neoplasias Colorretais/genética , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Inibidor p16 de Quinase Dependente de Ciclina/metabolismo , Inibidor p16 de Quinase Dependente de Ciclina/genética , Cobre/metabolismo , Regulação Neoplásica da Expressão Gênica , Transição Epitelial-Mesenquimal/genética , Linhagem Celular Tumoral , Resistencia a Medicamentos Antineoplásicos/genética , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismoRESUMO
The glial cell line-derived neurotrophic factor (GDNF) potential as a therapeutic agent for the treatment of Parkinson's Disease (PD) has been extensively explored. However, the mechanism of the GDNF neuroprotective effects is still unclear. In this study, the neuroprotective mechanism of the GDNF in the PD cellular models, which was obtained by the 6-hydroxydopamine (6-OHDA)-induced dopaminergic (DA) cell line MN9D damage was investigated by microarray. Interestingly, 54 constitutively increased or decreased genes were detected, 17 of which have not been reported previously. The expression of 5 up-regulated and 5 down-regulated genes which displayed the most obvious changes compared to the no GDNF treatment cells and was previously proven to be related to cell survival was validated by real-time PCR and western blot. Moreover, the up-regulated gene Ager and down-regulated gene Ccnl2 which were related to the PI-3K/Akt signaling pathway, but not researched in the neuron-cells, were investigated by overexpression and RNA interference. Overexpression of Ager or knockdown the expression of Ccnl2 decreased the damage to MN9D cells caused by 6-OHDA and reduced their apoptosis. All these results suggested that the protective effects of the GDNF on the 6-OHDA damaged MN9D cells could be understood by enhancing the expression of the apoptosis inhibiting genes and decreasing the expression of the apoptosis promoting genes. Thus, this study might provide a number of specific candidates and potential targets to investigate the protective mechanism of GDNF in DA neurons.
Assuntos
Fator Neurotrófico Derivado de Linhagem de Célula Glial/uso terapêutico , Modelos Biológicos , Fármacos Neuroprotetores/uso terapêutico , Doença de Parkinson/tratamento farmacológico , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Linhagem Celular , Análise por Conglomerados , Ciclinas/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Fator Neurotrófico Derivado de Linhagem de Célula Glial/farmacologia , Camundongos , Fármacos Neuroprotetores/farmacologia , Análise de Sequência com Séries de Oligonucleotídeos , Oxidopamina , Reação em Cadeia da Polimerase em Tempo Real , Receptor para Produtos Finais de Glicação Avançada , Receptores Imunológicos/metabolismo , Reprodutibilidade dos Testes , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genéticaRESUMO
Glial-cell-line-derived neurotrophic factor (GDNF) has been shown to protect dopaminergic (DA) neurons against 6-hydroxydopamine (6-OHDA) toxicity. The mechanism underlying the antiapoptosis role of GDNF still needs further studies. We previously observed that nuclear factor-kappaB (NF-κB) signaling pathway, i.e. p65/p52, mediated the antiapoptosis role of GDNF in MN9D cells. Here, the DA cell line MN9D was used to explore the mechanisms underlying NF-κB p65/p52-mediated protection role of GDNF in DA neurons. The results showed that GDNF pretreatment blocked the apoptotic effects induced by 6-OHDA, with the upregulation of the antiapoptotic protein, Bcl-2 and Bcl-w, as well as the downregulation of the proapoptotic proteins, Bax and Bad. Furthermore, when sip100 plasmids were transfected into MN9D cells to inhibit the expression of p100, which was the precursor of p52, the effects of GDNF on upregulating Bcl-2 and Bcl-w were attenuated. These results indicated that GDNF could protect MN9D cells from apoptosis induced by 6-OHDA via upregulating Bcl-2 and Bcl-w expressions and downregulating Bax and Bad expressions. Moreover, NF-κB p65/p52 signaling mediated the effects of GDNF on Bcl-2 and Bcl-w expressions.
Assuntos
Apoptose/efeitos dos fármacos , Neurônios Dopaminérgicos/metabolismo , Genes bcl-2/genética , Fator Neurotrófico Derivado de Linhagem de Célula Glial/genética , Hidroxidopaminas/farmacologia , Subunidade p52 de NF-kappa B/genética , Proteínas/genética , Fator de Transcrição RelA/genética , Animais , Proteínas Reguladoras de Apoptose , Western Blotting , Neurônios Dopaminérgicos/efeitos dos fármacos , Mesencéfalo/citologia , Camundongos , Subunidade p52 de NF-kappa B/fisiologia , Plasmídeos/genética , RNA Interferente Pequeno/genética , Fator de Transcrição RelA/fisiologia , Transfecção , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo , Proteína de Morte Celular Associada a bcl/genética , Proteína de Morte Celular Associada a bcl/metabolismoRESUMO
G protein-coupled receptors (GPCRs), the largest cell surface receptor superfamily, are involved in many physiological and pathological processes. G protein-coupled receptor 3 (Gpr3) is a newly discovered sphingosine 1-phosphate receptor, which directly or indirectly takes part in regulating the processes of nervous system and follicle development in the vertebrates. As a potential therapeutic drug target for a variety of neurological diseases and premature ovarian failure, its physiological function and biological mechanisms deserve further studies. In this paper, we reviewed the functions of Gpr3 in the processes of nervous system development and ovarian follicular development in the vertebrates.